Stable polymorph of N-(3-ethynylphenyl)-6, 7-bis (2-methoxyethoxy)-4-quinazolinamine hydrochloride, methods of production, and pharmaceutical uses thereof

ABSTRACT

The present invention relates to a stable crystalline form of N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine hydrochloride designated the B polymorph, its production in essentially pure form, and its use. The invention also relates to the pharmaceutical compositions containing the stable polymorph B form of N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine as hydrochloride, as well other forms of the compound, and to methods of treating hyperproliferative disorders, such as cancer, by administering the compound.

This application claims the benefit of U.S. Provisional Application No.60/206,420, filed May 23, 2000, U.S. Provisional Application No.60/193,191, filed Mar. 30, 2000, and U.S. Provisional Application No.60/164,907, filed Nov. 11, 1999, the contents of which are herebyincorporated by reference.

Throughout this application various publications are referenced. Thedisclosures of these publications in their entireties are herebyincorporated by reference into this application in order to more fullydescribe the state of the art to which this invention pertains.

BACKGROUND OF THE INVENTION

N-(3-ethynylphenyl)-6,7-bis (2-methoxyethoxy)-4-quinazolinamine, ineither its hydrochloride or mesylate forms, or in an anhydrous andhydrous form, is useful in the treatment of hyperproliferativedisorders, such as cancers, in mammals.

U.S. Pat. No. 5,747,498, issued May 5, 1998, which is incorporatedherein by reference in its entirety, refers, in Example 20, to[6,7-bis(2-methoxyethoxy)-quinazolin-4-yl]-(3-ethynylphenyl)aminehydrochloride, which, the patent discloses, is an inhibitor of the erbBfamily of oncogenic and protooncogenic protein tyrosine kinases, such asepidermal growth factor receptor (EGFR), and is therefore useful for thetreatment of proliferative disorders, such as cancers, in humans.

The mesylate form, described in PCT International Publication No. WO99/55683 (PCT/IB99/00612, filed Apr. 8, 1999), the entire disclosure ofwhich is incorporated herein by reference, and assigned to a commonassignee, and shown in formula 1 below:

is useful for the treatment of proliferative disorders, and morepreferred with parenteral methods of administration, as compared to thehydrochloride compound, i.e. with greater effectiveness in solution.

The mesylate compounds are more soluble in aqueous compositions than thehydrochloride compound, and thus the mesylate compounds are easilydelivered according to parenteral methods of administration. Thehydrochloride compound is however preferred with respect to solidadministration such as with tablets and oral administration.

SUMMARY OF THE INVENTION

The present invention relates to polymorphs, and methods for theselective production of polymorphs ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinaminehydrochloride, particularly in the stable polymorph form.

The present invention also relates to novel uses ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine, ineither its hydrochloride or mesylate forms, in an anhydrous or hydrousform, as well as in its various polymorph forms, in the treatment ofhyperproliferative disorders, such as cancers, in mammals.

DESCRIPTION OF THE FIGURES

FIG. 1 The X-ray powder diffraction patterns for the hydrochloridepolymorph A, the thermodynamically less stable form, over a larger rangeto show the first peaks.

FIG. 2 The X-ray powder diffraction patterns for the hydrochloridepolymorph A, the thermodynamically less stable form, are over a shorterrange to show more detail.

FIG. 3 The X-ray powder diffraction patterns for the hydrochloridepolymorph B, the thermodynamically more stable form, over a larger rangeto show the first peaks.

FIG. 4 The X-ray powder diffraction patterns for the hydrochloridepolymorph B, the thermodynamically more stable form, over a shorterrange to show more detail.

DETAILED DESCRIPTION OF THE INVENTION

Disclosed is a substantially homogeneous crystalline polymorph of thehydrochloride salt ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinaminedesignated the B polymorph that exhibits an X-ray powder diffractionpattern having characteristic peaks expressed in degrees 2-theta atapproximately 6.26, 12.48, 13.39, 16.96, 20.20, 21.10, 22.98, 24.46,25.14 and, 26.91. The polymorph is also characterized by the X-raypowder diffraction pattern shown in FIG. 3.

Disclosed is a crystalline polymorph of the hydrochloride salt ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinaminedesignated the B polymorph that exhibits an X-ray powder diffractionpattern having characteristic peaks expressed in degrees 2-theta atapproximately 6.26, 12.48, 13.39, 16.96, 20.20, 21.10, 22.98, 24.46,25.14 and, 26.91, which is substantially free of the polymorphdesignated the A polymorph. The polymorph is also characterized by theX-ray powder diffraction pattern shown in FIG. 3.

The polymorph designated the B polymorph may be in substantially pureform, relative to the A polymorph.

Also disclosed is a composition comprising a substantially homogeneouscrystalline polymorph of the hydrochloride salt ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine thatexhibits an X-ray powder diffraction pattern having characteristic peaksexpressed in degrees 2-theta at approximately 6.26, 12.48, 13.39, 16.96,20.20, 21.10, 22.98, 24.46, 25.14 and, 26.91. The hydrochloride salt ofN-(3-ethynylphenyl)-6,7-bis (2-methoxyethoxy)-4-quinazolinamine alsoexhibits an X-ray powder diffraction pattern having characteristic peaksexpressed in degrees 2-theta at approximately the values show in Table 3or in Table 4 below. And, theN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinaminehydrochloride in the polymorph B form may characterized by the X-raypowder diffraction pattern shown in FIG. 3.

Also disclosed is a composition comprising a crystalline polymorph ofthe hydrochloride salt ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinaminedesignated the B polymorph that exhibits an X-ray powder diffractionpattern having characteristic peaks expressed in degrees 2-theta atapproximately 6.26, 12.48, 13.39, 16.96, 20.20, 21.10, 22.98, 24.46,25.14 and, 26.91 in a weight % of the B polymorph relative to the Apolymorph which is at least 70%. This composition may comprise at least75% polymorph B, by weight; at least 80% polymorph B, by weight; atleast 85% polymorph B, by weight; at least 90% polymorph B, by weight;at least 95% polymorph B, by weight; at least 97% polymorph B, byweight; at least 98% polymorph B, by weight; or at least 99% polymorphB, by weight relative to the A polymorph.

Further disclosed is a process for producing the polymorph B of thehydrochloride salt ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine byrecrystallization ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinaminehydrochloride in a solvent comprising alcohol and water.

In the process, the recrystallization may comprise the steps of:

-   -   a) heating to reflux alcohol, water and the hydrochloride salt        of        N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine        so as to form a solution;    -   b) cooling the solution to between about 65 and 70° C.;    -   c) clarifying the solution; and    -   d) precipitating polymorph B by further cooling the clarified        solution.

In the process, theN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinaminehydrochloride is prepared by the steps of:

-   -   coupling a compound of formula 6        with a compound of formula 4

The compound of formula 6 is prepared by reacting a compound of formulaformula 5

in a suspension of metal alkali and solvent and with heating.

The compound of formula 4 is prepared by chlorinating a compound offormula 3

Also disclosed is a pharmaceutical composition for the treatment of ahyperproliferative disorder in a mammal which substantially comprises atherapeutically effective amount of the polymorph B and apharmaceutically acceptable carrier.

The pharmaceutical composition may be adapted for oral administration.It may be in the form of a tablet.

Also disclosed is a method of treating a hyperproliferative disorder ina mammal which comprises administering to said mammal a therapeuticallyeffective amount of the polymorph B.

The method may be for the treatment of a cancer selected from brain,squamous cell, bladder, gastric, pancreatic, breast, head, neck,oesophageal, prostate, colorectal, lung, renal, kidney, ovarian,gynecological and thyroid cancer.

The method may also be for the treatment of a cancer selected fromnon-small cell lung cancer (NSCLC), refractory ovarian cancer, head andneck cancer, colorectal cancer and renal cancer.

In the method, the therapeutically effective amount may be from about0.001 to about 100 mg/kg/day, or from about 1 to about 35 mg/kg/day.

In the method, the therapeutically effective amount may also be fromabout 1 to about 7000 mg/day; from about 5 to about 2500 mg/day; fromabout 5 to about 200 mg/day; or from about 25 to about 200 mg/day.

Further disclosed is a method for the treatment of a hyperproliferativedisorder in a mammal which comprises administering to said mammal atherapeutically effective amount of the polymorph B in combination withan anti-tumor agent selected from the group consisting of mitoticinhibitors, alkylating agents, anti-metabolites, intercalatingantibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes,topoisomerase inhibitors, biological response modifiers, anti-hormones,and anti-androgens.

Yet further disclosed is a method of making a composition whichcomposition comprises substantially homogeneous crystalline polymorph ofthe hydrochloride salt ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinaminedesignated the B polymorph that exhibits an X-ray powder diffractionpattern having characteristic peaks expressed in degrees 2-theta atapproximately 6.26, 12.48, 13.39, 16.96, 20.20, 21.10, 22.98, 24.46,25.14 and, 26.91, comprising admixing the crystalline polymorphdesiganted the B polymorph with a carrier.

The carrier may be a pharmaceutically acceptable carrier.

Also disclosed is a method of preparing polymorph B ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinaminehydrochloride which comprises the step of recrystallizingN-(3-ethynylphenyl)-6,7-bis (2-methoxyethoxy)-4-quinazolinaminehydrochloride in a solvent comprising alcohol.

In the method the solvent may further comprises water.

In the method, the N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine hydrochloride is prepared bycoupling a compound of formula 6

-   -   with a compound of formula 4

In the method, the compound of formula 6 is prepared by reacting acompound of formula 5

in a suspension of metal alkali and solvent and with heating.

In the method, the compound of formula 4 is prepared by chlorinating acompound of formula 3

Further disclosed is a method for the production of the polymorph B ofclaim 1 comprising the steps of:

-   -   a) substitution chlorination of starting quinazolinamine        compound of formula 3        having an hydroxyl group, to provide a compound of formula 4    -   by reaction thereof in a solvent mixture of thionyl chloride,        methylene chloride and dimethylformamide,    -   b) preparation of a compound of formula 6        in situ from starting material of compound of formula 5        by reaction of the latter in a suspension of metal alkali and        solvent and with heating;    -   c) reaction of the compound of formula 6 in situ with the        compound of formula 4 wherein the compound of formula 6 replaces        the chlorine in the compound of formula 4 to give the        N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine        hydrochloride;    -   d) recrystallizing the        N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine        hydrochloride, in alcohol, into the polymorph B form.

In this method, the substitution chlorination may be quenched in thepresence of aqueous sodium hydroxide; aqueous sodium bicarbonate;aqueous potassium hydroxide; aqueous potassium bicarbonate; aqueouspotassium carbonate; aqueous sodium carbonate, or a mixture thereof.

Yet further disclosed is a method for the production of polymorph B ofthe hydrochloride salt ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine byrecrystallization comprising the steps of:

-   -   a) heating to reflux alcohol, water and the hydrochloride salt        of N-(3-ethynylphenyl)-6,7-bis        (2-methoxyethoxy)-4-quinazolinamine so as to form a solution;    -   b) cooling the solution to between about 65 and 70° C.;    -   c) clarifying the solution; and    -   d) precipitating polymorph B by further cooling the clarified        solution.

Further disclosed is a composition consisting essentially ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinaminehydrochloride in the form of polymorph A, which is characterized by thefollowing peaks in its X-ray powder diffraction pattern shown in FIG. 1.

Also disclosed is a composition consisting essentially ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinaminehydrochloride in the form of polymorph A, which is characterized by thepeaks shown is Table 1 or Table 2 below.

A prodrug of any of the compound herein is also disclosed.

Further disclosed is a method of inducing differentiation of tumor cellsin a tumor comprising contacting the cells with an effective amount ofany of the compounds or compositions disclosed herein.

Also discosed is a method for the treatment of NSCLC (non small celllung cancer), pediatric malignancies, cervical and other tumors causedor promoted by human papilloma virus (HPV), melanoma, Barrett'sesophagus (pre-malignant syndrome), adrenal and skin cancers and autoimmune, neoplastic cutaneous diseases and atherosclerosis in a mammalcomprising administering to said mammal a therapeutically effectiveamount of a pharmaceutical composition comprised of at least one ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine, andpharmaceutically acceptable salts thereof in anhydrous and hydrateforms.

The treatment may further comprise a palliative or neo-adjuvant/adjuvantmonotherapy; or comprises blocking epidermal growth factor receptors(EGFR).

The method of may also be used in the treatment of tumors that expressEGFRvIII.

The method may further comprise a combination with any of chemotherapyand immunotherapy; or treatment with either or both anti-EGFR andanti-EGF antibodies; or administration to said mammal of a member of thegroup consisting of inhibitors of MMP (matrix-metallo-proteinase), VEGFR(vascular endothelial growth factor receptor), farnesyl transferase,CTLA₄. (cytotoxic T-lymphocyte antigen 4) and erbB2, MAb to VEGFr,rhuMAb-VEGF, erbB2 MAb and avb3 Mab.

The pharmaceutical compounds used may be radiation sensitizers forcancer treatment or in combination with anti-hormonal therapies, or forthe inhibition of tumor growth in humans in a regimen with radiationtreatment.

Further disclosed is a method for the chemoprevention of basal orsquamous cell carcinoma of the skin in areas exposed to the sun or inpersons of high risk to said carcinoma, said method comprisingadministering to said persons a therapeutically effective amount of apharmaceutical composition comprised of at least one ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine, andpharmaceutically acceptable salts thereof in anhydrous and hydrateforms.

Also is a method of inducing differentiation of tumor cells in a tumorcomprising contacting the cells with an effective amount of the compoundof at least one ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine, andpharmaceutically acceptable salts thereof in anhydrous and hydrateforms.

It is accordingly an object of the present invention to provide a methodfor the production of the hydrochloride salt ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine in HClform (Formula 2):

making it more suitable for tablet and oral administration andconsisting essentially of the stable polymorphic form (polymorph form B)as well as the compound in such polymorph B form and the intermediatepolymorph A in essentially pure form.

It is a further object of the present invention to provide such stablepolymorph form B in a pharmaceutical orally administered composition.

Stability of the hydrochloride compound is of concern for its use in thetreatment of patients since variations will affect effective dosagelevel and administration. It has been discovered that the hydrochlorideof N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine existsin two polymorph states, polymorph A and B. This contrasts with themesylate compounds which exist in three polymorph states (mesylatepolymorphs A, B and C). Polymorph B of the hydrochloride was found to bethe thermodynamically most stable and desirable form and the presentinvention comprises the polymorph B compound in the substantially purepolymorphic B form and pharmaceutical compositions of the substantiallypure form of polymorph B, particularly in tablet form and a method ofthe selective production of the compound.

The hydrochloride compound disclosed in the U.S. Pat. No. 5,747,498actually comprised a mixture of the polymorphs A and B, which, becauseof its partially reduced stability (i.e., from the polymorph Acomponent) was not more preferred for tablet form than the mesylate saltforms.

Specifically, the present invention relates to methods of producing thehydrochloride compound forms ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine and forproducing the stable form B in high yield. The mesylate salt ofN-(3-ethynylphenyl)-6,7-bis (2-methoxyethoxy)-4-quinazolinamine has beendiscovered to exist in at least three polymorphic forms which have beendesignated A, B, and C, of increasing stability with different X-raypowder diffraction patterns. The X-ray powder diffraction patterns forthe hydrochloride polymorph A (A₁ and A₂) and B (B₁ and B₂) forms areshown in FIGS. 1-4 as follows: graphs of FIGS. 1 and 3 are over a largerrange to fully show the first peaks for A and B, respectively, andgraphs of FIGS. 2 and 4 are over a shorter range to show more overalldetail for A and B, respectively.

The data contained in the above X-ray diffraction patterns of FIG. 1-4are tabulated in the following Tables 1-4: TABLE 1 Polymorph A Anode:Cu - Wavelength 1: 1.54056 Wavelength 2: 1.54439 (Rel Intensity: 0.500)Range# 1 - Coupled: 3.000 to 40.000 StepSize: 0.040 StepTime: 1.00Smoothing Width: 0.300 Threshold: 1.0 d(A) l(rel) 15.82794 100.014.32371 3.9 11.74376 1.5 11.03408 1.2 10.16026 1.4 8.98039 13.1 7.858257.8 6.63179 1.7 5.84901 2.1 5.69971 2.3 5.46922 2.4 5.21396 3.6 4.805693.5 4.70077 12.2 4.54453 4.8 4.19685 4.7 4.16411 4.4 3.97273 4.7 3.9134412.4 3.78223 24.2 3.67845 8.8 3.61674 8.2 3.50393 9.3 3.40200 6.03.35174 5.3 3.29005 4.2 3.05178 7.1 2.97750 3.0 2.91238 3.5 2.73148 3.72.60193 1.8 2.48243 1.3 2.40227 2.2 2.31297 1.7

TABLE 2 Polymorph A Anode: Cu - Wavelength 1: 1.54056 Wavelength 2:1-54439 (Rel Intensity: 0.500) Range#1 - Coupled: 3.000 to 40.000StepSize: 0.040 StepTime: 1.00 Smoothing Width: 0.300 Threshold: 1.02-Theta l(rel) 5.579 100.0 6.165 3.9 7.522 1.5 8.006 1.2 8.696 1.4 9.84113.1 11.251 7.8 13.340 1.7 15.135 2.1 15.534 2.3 16.193 2.4 16.991 3.618.447 3.5 18.862 12.2 19.517 4.8 21.152 4.7 21.320 4.4 22.360 4.722.703 12.4 23.502 24.2 24.175 8.8 24.594 8.2 25.398 9.3 26.173 6.026.572 5.3 27.080 4.2 29.240 7.1 30.007 3.0 30.673 3.5 32.759 3.7 34.4401.8 36.154 1.3 37.404 2.2 38.905 1.7

TABLE 3 Polymorph B Anode: Cu - Wavelength 1 1.54056 Wavelength 2:1.54439 (Rel Intensity: 0.500) Range # 1 - Coupled 3.000 to 40.040StepSize: 0.040 StepTime 1.00 Smoothing Width: 0.300 Threshold: 1.0 d(A)l(rel) 14.11826 100.0 11.23947 3.2 9.25019 3.9 7.74623 1.5 7.08519 6.46.60941 9.6 5.98828 2.1 5.63253 2.9 5.22369 5.5 5.01567 2.5 4.87215 0.74.72882 1.5 4.57666 1.0 4.39330 14.4 4.28038 4.2 4.20645 14.4 4.060074.7 3.95667 4.5 3.86656 4.8 3.76849 2.3 3.71927 3.0 3.63632 6.8 3.5396710.0 3.47448 3.7 3.43610 3.9 3.35732 2.8 3.31029 5.6 3.23688 0.9 3.167551.5 3.11673 4.3 3.07644 1.4 2.99596 2.1 2.95049 0.9 2.89151 1.6 2.839922.2 2.81037 2.4 2.74020 1.7 2.69265 1.7 2.58169 1.5 2.51043 0.8 2.473561.0 2.43974 0.6 2.41068 1.1 2.38755 1.4 2.35914 1.7

TABLE 4 Polymorph B Anode: Cu - Wavelength 1 1.54056 Wavelength 2:1.54439 (Rel Intensity: 0.500) Range# 1 - Coupled: 3.000 to 40.040StepSize 0.040 StepTime: 1.00 Smothing Width: 0.300 Threshold: 1.02-Theta l(rel) 6.255 100.0 7.860 3.2 9.553 3.9 11.414 1.5 12.483 6.413.385 9.6 14.781 2.1 15.720 2.9 16.959 5.5 17.668 2.5 18.193 0.7 18.7491.5 19.379 1.0 20.196 14.4 20.734 4.2 21.103 14.4 21.873 4.7 22.452 4.522.982 4.8 23.589 2.3 23.906 3.0 24.459 6.8 25.138 10.0 25.617 3.725.908 3.9 26.527 2.8 26.911 5.6 27.534 0.9 28.148 1.5 28.617 4.3 29.0001.4 29.797 2.1 30.267 0.9 30.900 1.6 31.475 2.2 31.815 2.4 32.652 1.733.245 1.7 34.719 1.5 35.737 0.8 36.288 1.0 36.809 0.6 37.269 1.1 37.6431.4 38.114 1.7

It is to be understood that the X-ray powder diffraction pattern is onlyone of many ways to characterize the arrangement of atoms comprising thecompound N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinaminehydrochloride, and that other methods well known in the art, e.g. singlecrystal X-ray diffraction, may be used to identify in a sample,composition or other preparation the presence of polymorph B of thehydrochloride salt ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine.

The present invention relates to a compound which is polymorph B of thehydrochloride salt ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine thatexhibits an X-ray powder diffraction pattern having characteristic peaksexpressed in degrees 2-theta at approximately 6.26, 12.48, 13.39, 16.96,20.20, 21.10, 22.98, 24.46, 25.14 and, 26.91. This invention alsorelates to a polymorph of the hydrochloride salt ofN-(3-ethynylphenyl)-6,7-bis (2-methoxyethoxy)-4-quinazolinamine thatexhibits an X-ray powder diffraction pattern having characteristic peaksexpressed in degrees 2-theta at approximately the values shown in Table4 above.

This invention also relates to a compound which is polymorph A of thehydrochloride salt ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine thatexhibits an X-ray powder diffraction pattern having characteristic peaksexpressed in degrees 2-theta at approximately 5.58, 9.84, 11.25, 18.86,22.70, 23.50, 24.18, 24.59, 25.40 and 29.24. This invention also relatesto a polymorph of the hydrochloride salt ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine thatexhibits an X-ray powder diffraction pattern having characteristic peaksexpressed in degrees 2-theta at approximately the values shown in Table2 above.

Method of Production

The polymorph B in substantially pure form ofN-(3-ethynylphenyl)-6,7-bis (2-methoxyethoxy)-4-quinazolinaminehydrochloride (compound of formula 1) is prepared, in accordance withthe method of the present invention, by the steps of;

-   1) substitution chlorination of starting quinazolinamine compound    (formula 3):    having an hydroxyl group, such as by reaction thereof in a solvent    mixture of thionyl chloride, methylene chloride, and    dimethylformamide, and finally quenching the reaction with an    aqueous solution of sodium hydroxide or sodium bicarbonate. The    compound of formula 4:    is produced in high yield with replacement of the hydroxyl group    with chlorine;-   2) preparation of compound of formula 6:    from starting material of formula 5:    by reaction of the latter in a suspension of NaOH (or KOH, or a    combination) in toluene with heating;-   3) reaction of the compound of formula 6 with the compound of    formula 4 of step 1 wherein the compound of formula 6 replaces the    chlorine to give the    N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine    hydrochloride (compound of formula 2) with a. 97% yield.-   4) recrystallization of the compound of formula 2 (comprising both    polymorph A and polymorph B) into the more stable polymorph B in a    solvent comprising alcohol (e.g. 2B-ethanol) and water, generally in    high yield, e.g., about 85%.

Accordingly, the present invention relates to a method of preparingpolymorph B ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinaminehydrochloride which comprises recrystallization ofN-(3-ethynylphenyl)-6,7-bis (2-methoxyethoxy)-4-quinazolinaminehydrochloride in a solvent comprising alcohol and water. In oneembodiment, the method comprises the steps of heating to reflux alcohol,water and the hydrochloride salt of N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine so as to form a solution; coolingthe solution to between about 65 and 70° C.; clarifying the solution;and precipitating polymorph B by further cooling the clarified solution.In an embodiment, the alcohol is ethanol. In a preferred embodiment, theratio of ethanol to water is about 4:1. It is to be expected that otherlower alcohols, e.g., C₁-C₄ alcohols, are also suitable forrecrystallization of polymorph B with adjustment of the alcohol to waterratio as needed. In another preferred embodiment, the compound to berecrystallized is present in an amount relative to the total volume ofsolvent at a weight to volume ratio of about 0.05. In an embodiment,N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinaminehydrochloride is prepared by coupling a compound of formula 6 with acompound of formula 4. In another embodiment, the compound of formula 6is prepared by reacting a compound of formula 5 in a suspension of metalalkali and solvent, with heating.

In an embodiment, the compound of formula 4 is prepared by chlorinatinga compound of formula 3 by reaction of the latter in a solvent mixtureof thionyl chloride, methylene chloride and dimethylformide, andsubsequently quenching the reaction with an aqueous solution of sodiumhydroxide. Alternatively, an aqueous solution of sodium bicarbonate canbe substituted for the sodium hydroxide solution.

This invention relates to polymorph B of the hydrochloride salt ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine preparedby the above methods. In an embodiment, the polymorph B is prepared byusing the starting materials described herein. In a preferredembodiment, polymorph B is prepared by reaction of the startingmaterials described herein with the reagents and conditions according tothe methods described herein and in the Examples which follow.

General Synthesis

N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinaminehydrochloride has been found to exist in two distinct anhydrouspolymorphic forms A and B. The production method for the variouspolymorphs is with components separately reacted in accordance with thefollowing scheme:

Uses

As described in the aforementioned U.S. Pat. No. 5,747,498 and PCTInternational Publication No. WO 99/55683, the compounds made inaccordance with the present invention are useful for the treatment of ahyperproliferative disorder in a mammal which comprises atherapeutically effective amount of the hydrochloride or mesylate formof N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine, and apharmaceutically acceptable carrier.

The term “compound(s) of the invention” referred to herein is preferablythe polymorph B form of the hydrochloride salt ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinaminehydrochloride, but is not meant to exclude the mesylate form and itsthree polymorphs, or polymorph A of the hydrochloride form, or a mixtureof polymorphs B and A of the hydrochloride form or other non-crystallineforms of the compound.

The term “treating” as used herein, unless otherwise indicated, meansreversing, alleviating, inhibiting the progress of, or preventing thedisorder or condition to which such term applies, or one or moresymptoms of such disorder or condition. The term “treatment”, as usedherein, refers to the act of treating, as “treating” is definedimmediately above.

“Abnormal cell growth”, as used herein, refers to cell growth that isindependent of normal regulatory mechanisms (e.g., loss of contactinhibition), including the abnormal growth of normal cells and thegrowth of abnormal cells. This includes, but is not limited to, theabnormal growth of: (1) tumor cells (tumors), both benign and malignant,expressing an activated Ras oncogene; (2) tumor cells, both benign andmalignant, in which the Ras protein is activated as a result ofoncogenic mutation in another gene; (3) benign and malignant cells ofother proliferative diseases in which aberrant Ras activation occurs.Examples of such benign proliferative diseases are psoriasis, benignprostatic hypertrophy, human papilloma virus (HPV), and restenosis.“Abnormal cell growth” as used herein also refers to and includes theabnormal growth of cells, both benign and malignant, resulting fromactivity of the enzymes farnesyl protein transferase, protein kinases,protein phosphatases, lipid kinases, lipid phosphatases, or activity ortrascription factors, or intracellular or cell surface receptorproteins.

[6,7-bis(2-methoxyethoxy)quinazolin-4-yl]-(3-ethynylphenyl)-aminehydrochloride, preferably the stable polymorph B form, is additionallyused for the treatment of a variety of additional human tumorscontaining hyperproliferating cells that are activated by the signaltransduction pathways stimulated by EGFR, whether by overexpression(e.g. due to one or more of—altered transcription, altered mRNAdegradation or gene amplification) of the EGFR protein itself, anotherreceptor protein with which EGFR can form active heterodimers, or one ofthe ligands that activate EGFR (e.g. EGF, TGFα, amphiregulin,β-cellulin, heparin-binding EGF, or epiregulin) or a heterodimerizingreceptor, or due to a dependence or partial dependence on the activityof a “normal” level of EGFR protein, whether activated by extracellularligand, intracellular signal transduction pathways and/or geneticalterations or polymorphisms that result in amino acid substitutionsthat produce increased or ligand-independent activity (e.g. EGFRvIII,Archer G. E. et. al. (1999) Clinical Cancer Research 5:2646-2652). Suchtumors, including both benign and malignant, include renal (such askidney, renal cell carcinoma, or carcinoma of the renal pelvis), liver,kidney, bladder (particularly invasive tumors), breast (includingestrogen receptor negative and positive tumors, and progesteronereceptor negative and positive tumors), gastric, esophageal (includingBarrett's mucosa, squamous cell carcinomas and adenocarcinomas), larynx,ovarian, colorectal (particularly deeply invasive tumors), includinganal, prostate, pancreatic, lung (particularly non-small cell lungcancer (NSCLC) adenocarcinomas, large cell tumors and squamous cellcarcinomas, but also reactive (squamous metaplasia and inflammatoryatypia) as well as precancerous (dysplasia and carcinoma in situ)bronchial lesions associated with both NSCLC adenocarcinomas andsquamous cell carcinomas), gynecological, including vulval, endometrial,uterine (e.g, sarcomas), cervical, vaginal, vulval, and fallopian tubecancers, thyroid, hepatic carcinomas, skin cancers, sarcomas, braintumors, including glioblastomas (including gliobastoma multiforme),astrocytomas, schwanomas, ependymonas, medulloblastomas, meningiomas andpituitary adenomas, and various other head and neck tumors (particularlysquamous cell carcinomas), and metastases of all of the above.

[6,7-bis(2-methoxyethoxy)quinazolin-4-yl]-(3-ethynylphenyl)-aminehydrochloride, preferably the stable polymorph B form, is also used forthe treatment of a variety of additional human hyperplastic conditionscontaining hyperproliferating cells that are activated by the signaltransduction pathways capable of stimulation by EGFR, such as benignhyperplasia of the skin (e.g. psoriasis) or prostate (e.g. BPH), chronicpancreatitis, or reactive hyperplasia of pancreatic ductal epithelium,or kidney disease (including proliferative glomerulonephritis anddiabetes-induced renal disease) in a mammal which composition comprisesa therapeutically effective amount of the hydrochloride ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine,preferably the polymorph B form, and a pharmaceutically acceptablecarrier.

In addition, pharmaceutical compositions including the compounds made inaccordance with the present invention provide for the prevention ofblastocyte implantation in a mammal, which composition comprises atherapeutically effective amount ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinaminehydrochloride, preferably the polymorph B form, and a pharmaceuticallyacceptable carrier.

[6,7-bis (2-methoxyethoxy) quinazolin-4-yl]-(3-ethynylphenyl)-aminehydrochloride, preferably the stable polymorph B form, is also used forthe treatment of additional disorders in which cells are activated bythe signal transduction pathways stimulated by EGFR, whether byoverexpression (due to one or more of—altered transcription, alteredmRNA degradation or gene amplification) of the EGFR protein itself,another receptor protein with which it can form active heterodimers, orone of the ligands that activate EGFR (e.g. EGF, TGFα, amphiregulin,β-cellulin, heparin-binding EGF, or epiregulin) or a heterodimerizingreceptor, or due to a dependence or partial dependence on the activityof a “normal” level of EGFR protein, whether activated by extracellularligand, intracellular signal transduction pathways and/or geneticalterations or polymorphisms that result in amino acid substitutionsthat produce increased or ligand-independent activity (e.g. EGFRvIII,Archer G. E. et. al. (1999) Clinical Cancer Research 5:2646-2652). Suchdisorders may include those of a neuronal, glial, astrocytal,hypothalamic, and other glandular, macrophagal, epithelial, stromal, orblastocoelic nature in which aberrant or ‘normal’ function, expression,activation or signalling via EGFR may be involved. Such disorders mayfurthermore involve the modulation by EGF (or other ligands thatactivate EGFR or heterodimerizing receptors) of adipocyte lipogenesis,bone resorption, hypothalamic CRH release, hepatic fat accumulation,T-cell proliferation, skin tissue proliferation or differentiation,corneal epithelial tissue proliferation or differentiation, macrophagechemotaxis or phagocytosis, astroglial proliferation, wound healing,polycystic kidney disease, lung epithelial proliferation ordifferentiation (e.g. associated with asthmatic airway remodeling ortissue repair), inflammatory arthritis (e.g. rheumatoid arthritis,systemic lupus erythematosus-associated arthritis, psoriatis arthritis)testicular androgen production, thymic epithelial cell proliferation,uterine epithelial cell proliferation, angiogenesis, cell survival,apoptosis, NFκB activation, vascular smooth muscle cell proliferation,restenosis or lung liquid secretion.

[6,7-bis(2-methoxyethoxy)quinazolin-4-yl]-(3-ethynylphenyl)-aminehydrochloride, preferably the stable polymorph B form, is also used forthe treatment of a range of leukemias (chronic and acute) and lymphoidmalignancies (e.g. lymphocytic lymphomas), diabetes, diabetic and otherretinopathies, such as retinophay or prematurity, age-related maculardegeneration, solid tumors of childhood, glioma, hemangiomas, melanomas,including intraocular or uveal melanomas, Kaposi's sarcoma, Hodgkin'sdisease, epidermoid cancers, cancers of the endocrine system (e.g.parathyroid, adrenal glands), bone small intestine, urethra, penis andureter, atherosclerosis, skin diseases such as eczema and scleroderma,mycoses fungoides, sarcomas of the soft tissues and neoplasm of thecentral nervous system (e.g. primary CNS lymphoma, spinal axis tumors,brain stem gliomas, or pituitary adenomas).

The treatment of any of the hyperproliferative or additional disordersdescribed above may be applied as a monotherapy, or may involve inaddition to[6,7-bis(2-methoxyethoxy)quinazolin-4-yl]-(3-ethynylphenyl)-aminehydrochloride, preferably the stable polymorph B form, application withone or more additional drugs or treatments (e.g. radiotherapy,chemoradiotherapy) that are anti-hyperproliferative, anti-tumor orantihyperplastic in nature. Such conjoint treatment may be achieved byway of simultaneous, sequential, cyclic or separate dosing of theindividual components of the treatment.[6,7-bis(2-methoxyethoxy)quinazolin-4-yl]-(3-ethynylphenyl)-aminehydrochloride, preferably the stable polymorph B form, is typically usedat doses of 1-7000 mg/day, preferably 5-2500 mg/day, most preferably5-200 mg/day, for any of the above treatments.

Furthermore, the various forms ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine includingthe mesylate and hydrochloride forms (all polymorph forms) as well asother pharmaceutically acceptable salt forms, and anhydrous and hydrateforms, can be used for treatment, with a therapeutically-effectiveamount of the aforementioned compounds and a pharmaceutically acceptablecarrier, of the specific conditions of NSCLC (non small cell lungcancer), pediatric malignancies, cervical and other tumors caused orpromoted by human papilloma virus (HPV), melanoma, Barrett's esophagus(pre-malignant syndrome) and adrenal and skin cancers as well as autoimmune and neoplastic cutaneous diseases such as mycoses fungoides, in amammal, as well as for the chemoprevention of basal or squamous cellcarcinomas of the skin, especially in areas exposed to the sun or inpersons known to be at high risk for such cancers. In addition, theaforementioned compounds are useful in treatment of atherosclerosis,with epidermal growth factor having been implicated in thehyperproliferation of vascular smooth muscle cells responsible foratherosclerotic plaques (G. E. Peoples et al., Proc. Nat. Acad. Sci. USA92:6547-6551, 1995).

The compounds of the present invention are potent inhibitors of the erbBfamily of oncogenic and protooncogenic protein tyrosine kinases such asepidermal growth factor receptor (EGFR), erbB2, HER3, or HER4 and thusare all adapted to therapeutic use as antiproliferative agents (e.g.,anticancer) in mammals, particularly in humans. The compounds of thepresent invention are also inhibitors of angiogenesis and/orvasculogenesis.

The compounds of the present invention may also be useful in thetreatment of additional disorders in which aberrant expressionligand/receptor interactions or activation or signalling events relatedto various protein tyrosine kinases are involved. Such disorders mayinclude those of neuronal, glial, astrocytal, hypothalamic, glandular,macrophagal, epithelial, stromal, or blastocoelic nature in whichaberrant function, expression, activation or signalling of the erbBtyrosine kinases are involved. In addition, the compounds of the presentinvention may have therapeutic utility in inflammatory, angiogenic andimmunologic disorders involving both identified and as yet unidentifiedtyrosine kinases that are inhibited by the compounds of the presentinvention.

In addition to direct treatment of the above ailments with thecompounds, the utilization and treatment in these and generalapplications may be as palliative or neo-adjuvant/adjuvant monotherapy,in blocking epidermal growth factor receptors (EGFR) and for use intreatment of tumors that express a variant form of EGFR known asEGFRvIII as described in the scientific literature (e.g., D K Moscatelloet al. Cancer Res. 55:5536-5539, 1995), as well as in a combination withchemotherapy and immunotherapy. As described in more detail below,treatment is also possible with both anti-EGFR and anti-EGF antibodycombinations or with combination of inhibitors of MMP(matrix-metallo-proteinase), other tyrosine kinases including VEGFR(vascular endothelial growth factor receptor), farnesyl transferase,CTLA₄. (cytotoxic T-lymphocyte antigen 4) and erbB2. Further treatmentsinclude MAb to VEGFr, and other cancer-related antibodies includingrhuMAb-VEGF (Genentech, Phase III), the erbB2 MAb available as Herceptin(Genentech, Phase III), or the avb3 MAb available as Vitaxin (AppliedMolecular Evolution/MedImmune, Phase II).

The invention also relates to a pharmaceutical composition and a methodof treating any of the mentioned disorders in a mammal which comprisesadministering to said mammal a therapeutically effective amount ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine,preferably in hydrochloride polymorph B form, and a pharmaceuticallyacceptable carrier.

Combination Therapy

The active compound may be applied as a sole therapy or may involve oneor more other materials and treatment agents such as both anti-EGFR andanti-EGF antibody combinations or with combination of inhibitors of MMP(matrix-metallo-proteinase), other tyrosine kinases including VEGFR(vascular endothelial growth factor receptor), farnesyl transferase,CTLA₄. (cytotoxic T-lymphocyte antigen 4) and erbB2, as well as MAb toVEGFr, and other cancer-related antibodies including rhuMAb-VEGF, theerbB2 MAb, or avb3.

Thus, the active compound may be applied with one or more otheranti-tumor substances, for example those selected from, for example,mitotic inhibitors, for example vinblastine; alkylating agents, forexample cis-platin, carboplatin and cyclophosphamide; anti-metabolites,for example 5-fluorouracil, cytosine arabinoside and hydroxyurea, or,for example, one of the preferred anti-metabolites disclosed in EuropeanPatent Application No. 239362 such asN-(5-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]-2-thenoyl)-L-glutamicacid; growth factor inhibitors; cell cycle inhibitors; intercalatingantibiotics, for example adriamycin and bleomycin; enzymes, for exampleinterferon; and anti-hormones, for example anti-estrogens such asNolvadex® (tamoxifen) or, for example anti-androgens such as Casodex®(4′-cyano-3-(4-fluorophenylsulphonyl)-2-hydroxy-2-methyl-3′-(trifluoromethyl)propionanilide).

In a further embodiment, the compounds of the invention may beadministered in conjunction with an anti-angiogenesis agent(s) such as aMMP-2 (matrix-metalloproteinase-2) inhibitor(s), a MMP-9(matrix-metalloproteinase-9) inhibitor(s), and/or COX-II (cyclooxygenaseII) inhibitor(s) in the methods of treatment an compositions describedherein. For the combination therapies and pharmaceutical compositionsdescribed herein, the effective amounts of the compound of the inventionand of the chemotherapeutic or other agent useful for inhibitingabnormal cell growth (e.g., other antiproliferative agent,anti-angiogenic, signal transduction inhibitor or immune-systemenhancer) can be determined by those of ordinary skill in the art, basedon the effective amounts for the compound described herein and thoseknown or described for the chemotherapeutic or other agent. Theformulations and routes of administration for such therapies andcompositions can be based on the information described herein forcompositions and therapies comprising the compound of the invention asthe sole active agent and on information provided for thechemotherapeutic or other agent in combination therewith.

The invention also relates to production of compounds used in a methodfor the treatment of a hyperproliferative disorder in a mammal whichcomprises administering to said mammal a therapeutically effectiveamount of N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinaminehydrochloride in combination with an anti-tumor agent selected from thegroup consisting of mitotic inhibitors, alkylating agents,anti-metabolites, intercalating antibiotics, growth factor inhibitors,cell cycle inhibitors, enzymes, topoisomerase inhibitors, bioloicalresponse modifiers, anti-hormones, and anti-androgens.

The compounds are also useful as radiation sensitizers for cancertreatment and may be combined with anti-hormonal therapies. Parametersof adjuvant radiation therapies are for example contained inPCT/US99/10741, as published on 25 Nov. 1999, in InternationalPublication No. WO 99/60023, the disclosure of which is included hereinby reference thereto. With such mode of treatment for example, forinhibiting tumor growth, a radiation dosage of 1-100 Gy is utilizedpreferably in conjunction with at least 50 mg of the pharmaceuticalcompound, in a preferred dosage regimen of at least five days a week forabout two to ten weeks. Thus, this invention further relates to a methodfor inhibiting abnormal cell growth in a mammal which method comprisesadministering to the mammal an amount of the compound of the invention,or a pharmaceutically acceptable salt or solvate or prodrug thereof, incombination with radiation therapy, wherein the amount of the compound,salt, solvate or prodrug is in combination with the radiation therapyeffective in inhibiting abnormal cell growth in the mammal. Techniquesfor administering radiation therapy are known in the art, and thesetechniques can be used in the combination therapy described herein.

Anti-angiogenesis agents, such as MMP-2 (matrix-metalloproteinase 2)inhibitors, MMP-9 (matrix-metalloproteinase 9) inhibitors, and COX-II(cyclooxygenase II) inhibitors, can be used in conjunction with thecompound of the invention in the methods and pharmaceutical compositionsdescribed herein. Examples of useful COX-II inhibitors include CELEBREX™(alecoxib), valdecoxib, and rofecoxib. Examples of useful matrixmetalloproteinase inhibitors are described in WO 96/33172 (publishedOct. 24, 1996), WO 96/27583 (published Mar. 7, 1996), European PatentApplication No. 97304971.1 (filed Jul. 8, 1997), European PatentApplication No. 99308617.2 (filed Oct. 29, 1999), WO 98/07697 (publishedFeb. 26, 1998), WO 98/03516 (published Jan. 29, 1998), WO 98/34918(published Aug. 13, 1998), WO 98/34915 (published Aug. 13, 1998), WO98/33768 (published Aug. 6, 1998), WO 98/30566 (published Jul. 16,1998), European Patent Publication 606,046 (published Jul. 13, 1994),European Patent Publication 931,788 (published Jul. 28, 1999), WO90/05719 (published May 331, 1990), WO 99/52910 (published Oct. 21,1999), WO 99/52889 (published Oct. 21, 1999), WO 99/29667 (publishedJun. 17, 1999), PCT International Application No. PCT/IB98/01113 (filedJul. 21, 1998), European Patent Application No. 99302232.1 (filed Mar.25, 1999), Great Britain patent application number 9912961.1 (filed Jun.3, 1999), U.S. Provisional Application No. 60/148,464 (filed Aug. 12,1999), U.S. Pat. No. 5,863,949 (issued Jan. 26, 1999), U.S. Pat. No.5,861,510 (issued Jan. 19, 1999), and European Patent Publication780,386 (published Jun. 25, 1997), all of which are incorporated hereinin their entireties by reference. Preferred MMP-2 and MMP-9 inhibitorsare those that have little or no activity inhibiting MMP-1. Morepreferred, are those that selectively inhibit MMP-2 and/or MMP-9relative to the other matrix-metalloproteinases (i.e. MMP-1, MMP-3,MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12, and MMP-13).

Some specific examples of MMP inhibitors useful in the present inventionare AG-3340, RO 32-3555, RS 13-0830, and the compounds recited in thefollowing list:

-   0.3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-cyclopentyl)-amino]-propionic    acid;-   3-exo-3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]octane-3-carboxylic    acid hydroxyamide;-   (2R, 3R)    1-[4-(2-chloro-4-fluoro-benzyloxy)-benzenesulfonyl]-3-hydroxy-3-methyl-piperidine-2-carboxylic    acid hydroxyamide;-   4-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-4-carboxylic    acid hydroxyamide;-   3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-cyclobutyl)-amino]-propionic    acid;-   4-[4-(4-chloro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-4-carboxylic    acid hydroxyamide;-   (R)    3-[4-(4-chloro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-3-carboxylic    acid hydroxyamide;-   (2R, 3R)    1-[4-(4-fluoro-2-methyl-benzyloxy)-benzenesulfonyl]-3-hydroxy-3-methyl-piperidine-2-carboxylic    acid hydroxyamide;-   3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-1-methyl-ethyl)-amino]-propionic    acid;-   3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(4-hydroxycarbamoyl-tetrahydro-pyran-4-yl)-amino]-propionic    acid;-   3-exo-3-[4-(4-chloro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]octane-3-carboxylic    acid hydroxyamide;-   3-endo-3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]octane-3-carboxylic    acid hydroxyamide; and-   (R)    3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-tetrahydro-furan-3-carboxylic    acid hydroxyamide;    and pharmaceutically acceptable salts and solvates of said    compounds.

Other anti-angiogenesis agents, including other COX-II inhibitors andother MMP inhibitors, can also be used in the present invention.

The compound of the present invention can also be used with signaltransduction inhibitors, such as other agents that can inhibit EGFR(epidermal growth factor receptor) responses, such as EGFR antibodies,EGF antibodies, and other molecules that are EGFR inhibitors; VEGF(vascular endothelial growth factor) inhibitors, such as VEGF receptorsand molecules that can inhibit VEGF; and erbB2 receptor inhibitors, suchas other organic molecules or antibodies that bind to the erbB2receptor, for example, HERCEPTIN™ (Genentech, Inc. of South SanFrancisco, Calif., USA).

EGFR inhibitors are described in, for example in WO 95/19970 (publishedJul. 27, 1995), WO 98/14451 (published Apr. 9, 1998), WO 98/02434(published Jan. 22, 1998), and other compounds described in U.S. Pat.No. 5,747,498 (issued May 5, 1998), and such substances can be used inthe present invention as described herein. EGFR-inhibiting agentsinclude, but are not limited to, the monoclonal antibodies C225 andanti-EGFR 22Mab (ImClone Systems Incorporated of New York, N.Y., USA),the compounds ZD-1839 (AstraZeneca), BIBX-1382 (Boehringer Ingelheim),MDX-447 (Medarex Inc. of Annandale, N.J., USA), and OLX-103 (Merck & Co.of Whitehouse Station, N.J., USA), VRCTC-310 (Ventech Research) and EGFfusion toxin (Seragen Inc. of Hopkinton, Mass.). These and otherEGFR-inhibiting agents can be used in the present invention.

VEGF inhibitors, for example SU-5416 and SU-6668 (Sugen Inc. of SouthSan Francisco, Calif., USA), can also be combined with the compound ofthe present invention. VEGF inhibitors are described in, for example inWO 99/24440 (published May 20, 1999), PCT International ApplicationPCT/IB99/00797 (filed May 3, 1999), in WO 95/21613 (published Aug. 17,1995), WO 99/61422 (published Dec. 2, 1999), U.S. Pat. No. 5,834,504(issued Nov. 10, 1998), WO 98/50356 (published Nov. 12, 1998), U.S. Pat.No. 5,883,113 (issued Mar. 16, 1999), U.S. Pat. No. 5,886,020 (issuedMar. 23, 1999), U.S. Pat. No. 5,792,783 (issued Aug. 11, 1998), WO99/10349 (published Mar. 4, 1999), WO 97/32856 (published Sep. 12,1997), WO 97/22596 (published Jun. 26, 1997), WO 98/54093 (publishedDec. 3, 1998), WO 98/02438 (published Jan. 22, 1998), WO 99/16755(published Apr. 8, 1999), and WO 98/02437 (published Jan. 22, 1998), allof which are incorporated herein in their entireties by reference. Otherexamples of some specific VEGF inhibitors useful in the presentinvention are IM862 (Cytran Inc. of Kirkland, Wash., USA); anti-VEGFmonoclonal antibody of Genentech, Inc. of South San Francisco, Calif.;and angiozyme, a synthetic ribozyme from Ribozyme (Boulder, Colo.) andChiron (Emeryville, Calif.). These and other VEGF inhibitors can be usedin the present invention as described herein.

ErbB2 receptor inhibitors, such as GW-282974 (Glaxo Wellcome plc), andthe monoclonal antibodies AR-209 (Aronex Pharmaceuticals Inc. of TheWoodlands, Tex., USA) and 2B-1 (Chiron), can furthermore be combinedwith the compound of the invention, for example those indicated in WO98/02434 (published Jan. 22, 1998), WO 99/35146 (published Jul. 15,1999), WO 99/35132 (published Jul. 15, 1999), WO 98/02437 (publishedJan. 22, 1998), WO 97/13760 (published Apr. 17, 1997), WO 95/19970(published Jul. 27, 1995), U.S. Pat. No. 5,587,458 (issued Dec. 24,1996), and U.S. Pat. No. 5,877,305 (issued Mar. 2, 1999), which are allhereby incorporated herein in their entireties by reference. ErbB2receptor inhibitors useful in the present invention are also describedin U.S. Provisional Application No. 60/117,341, filed Jan. 27, 1999, andin U.S. Provisional Application No. 60/117,346, filed Jan. 27, 1999,both of which are incorporated in their entireties herein by reference.The erbB2 receptor inhibitor compounds and substance described in theaforementioned PCT applications, U.S. patents, and U.S. provisionalapplications, as well as other compounds and substances that inhibit theerbB2 receptor, can be used with the compound of the present inventionin accordance with the present invention.

The compound of the invention can also be used with other agents usefulin treating abnormal cell growth or cancer, including, but not limitedto, agents capable of enhancing antitumor immune responses, such asCTLA4 (cytotoxic lymphocyte antigen 4) antibodies, and other agentscapable of blocking CTLA4; and anti-proliferative agents such asfarnesyl protein transferase inhibitors. Specific CTLA4 antibodies thatcan be used in the present invention include those described in U.S.Provisional Application 60/113,647 (filed Dec. 23, 1998) which isincorporated by reference in its entirety, however other CTLA4antibodies can be used in the present invention.

Such conjoint treatment may be achieved by way of the simultaneous,sequential or separate dosing of the individual components of thetreatment.

It is expected that the compound of the invention can render abnormalcells more sensitive to treatment with radiation for purposes of killingand/or inhibiting the growth of such cells. Accordingly, this inventionfurther relates to a method for sensitizing abnormal cells in a mammalto treatment with radiation which comprises administering to the mammalan amount of the compound of the invention, pharmaceutically acceptablesalt or solvate thereof, or prodrug thereof, which amount is effectivein sensitizing abnormal cells to treatment with radiation. The amount ofthe compound, salt, solvate, or prodrug in this method can be determinedaccording to the means for ascertaining effective amounts of thecompound of the invention described herein.

The subject invention also includes isotopically-labelled compounds,which compounds are identical to the above recited compound of theinvention, but for the fact that one or more atoms thereof are replacedby an atom having an atomic mass or mass number different from theatomic mass or mass number usually found in nature. Examples of isotopesthat can be incorporated into the compound of the invention includeisotopes of hydrogen, carbon, nitrogen and oxygen, such as ²H, ³H, ¹³C,¹⁴C, ¹⁵N, ¹⁸O and ¹⁷O, respectively. Compounds of the present invention,and pharmaceutically acceptable salts of said compounds which containthe aforementioned isotopes and/or other isotopes of other atoms arewithin the scope of this invention. Certain isotopically-labelledcompounds of the present invention, for example those into whichradioactive isotopes such as ³H and ¹⁴C are incorporated, are useful indrug and/or substrate tissue distribution assays. Tritiated, i.e., ³H,and carbon-14, i.e., ¹⁴C, isotopes are particularly preferred for theirease of preparation and detectability. Further, substitution withheavier isotopes such as deuterium, i.e., ²H, can afford certaintherapeutic advantages resulting from greater metabolic stability, forexample increased in vivo half-life or reduced dosage requirements and,hence, may be preferred in some circumstances. Isotopically labelledcompounds of this invention can generally be prepared by carrying outthe procedures disclosed in the Methods and/or the examples below, andsubstituting a readily available isotopically labelled reagent for anon-isotopically labelled reagent, using methods well known in the art.Accordingly, reference to the compound of the invention for use in thetherapeutic methods and pharmaceutical compositions described hereinalso encompasses isotropically-labelled forms of the compound.

[6,7-bis(2-methoxyethoxy)quinazolin-4-yl]-(3-ethynylphenyl)-aminehydrochloride, preferably the stable polymorph B form, is typically usedat doses of 1-7000 mg/day, preferably 5-2500 mg/day, most preferably5-200 mg/day, for any of the above treatments.

Patients that can be treated with the compound of the invention, aloneor in combination, include, for example, patients that have beendiagnosed as having psoriasis, BPH, lung cancer, bone cancer, pancreaticcancer, skin cancer, cancer of the head and neck, cutaneous orintraocular melanoma, ovarian cancer, rectal cancer, cancer of the analregion, stomach cancer, colon cancer, breast cancer, gynecologic tumors(e.g., uterine sarcomas, carcinoma of the fallopian tubes, carcinoma ofthe endometrium, carcinoma of the cervix, carcinoma of the vagina orcarcinoma of the vulva), Hodgkin's disease, cancer of the esophagus,cancer of the small intestine, cancer of the endocrine system (e.g.,cancer of the thyroid, parathyroid or adrenal glands), sarcomas of softtissues, cancer of the urethra, cancer of the penis, prostate cancer,chronic or acute leukemia, solid tumors of childhood, lymphocyticlymphonas, cancer of the bladder, cancer of the kidney or ureter (e.g.,renal cell carcinoma, carcinoma of the renal pelvis), or neoplasms ofthe central nervous system (e.g., primary CNS lymphona, spinal axistumors, brain stem gliomas or pituitary adenomas).

Activity

The in vitro activity of the compounds of the present invention ininhibiting the receptor tyrosine kinase (and thus subsequentproliferative response, e.g., cancer) may be determined by the followingprocedure.

The activity of the compounds of the present invention, in vitro, can bedetermined by the amount of inhibition of the phosphorylation of anexogenous substrate (e.g., Lys₃-Gastrin or polyGluTyr (4:1) randomcopolymer (I. Posner et al., J. Biol. Chem. 267 (29), 20638-47 (1992))on tyrosine by epidermal growth factor receptor kinase by a testcompound relative to a control. Affinity purified, soluble human EGFreceptor (96 ng) is obtained according to the procedure in G. N. Gill,W. Weber, Methods in Enzymology 146, 82-88 (1987) from A431 cells(American Type Culture Collection, Rockville, Md.) and preincubated in amicrofuge tube with EGF (2 μg/ml) in phosphorylation buffer+vanadate(PBV: 50 mM HEPES, pH 7.4; 125 mM NaCl; 24 mM MgCl₂; 100 μM sodiumorthovanadate), in a total volume of 10 μl, for 20-30 minutes at roomtemperature. The test compound, dissolved in dimethylsulfoxide (DMSO),is diluted in PBV, and 10 μl is mixed with the EGF receptor/EGF mix, andincubated for 10-30 minutes at 30° C. The phosphorylation reaction isinitiated by addition of 20 μl ³³P-ATP/substrate mix (120 μMLys₃-Gastrin (sequence in single letter code for amino acids,KKKGPWLEEEEEAYGWLDF), 50 mM Hepes pH 7.4, 40 μM ATP, 2 μCi γ-[³³P]-ATP)to the EGFr/EGF mix and incubated for 20 minutes at room temperature.The reaction is stopped by addition of 10 μl stop solution (0.5 M EDTA,pH 8; 2 mM ATP) and 6 μl 2N HCl. The tubes are centrifuged at 14,000RPM, 4° C., for 10 minutes. 35 μl of supernatant from each tube ispipetted onto a 2.5 cm circle of Whatman P81 paper, bulk washed fourtimes in 5% acetic acid, 1 liter per wash, and then air dried. Thisresults in the binding of substrate to the paper with loss of free ATPon washing. The [³³P] incorporated is measured by liquid scintillationcounting. Incorporation in the absence of substrate (e.g., lys₃-gastrin)is subtracted from all values as a background and percent inhibition iscalculated relative to controls without test compound present. Suchassays, carried out with a range of doses of test compounds, allow thedetermination of an approximate IC₅₀ value for the in vitro inhibitionof EGFR kinase activity.

Other methods for determining the activity of the compounds of thepresent invention are described in U.S. Pat. No. 5,747,498, thedisclosure of which is incorporated herein.

Pharmaceutical Compositions

The pharmaceutical composition may, for example and most preferably, bein a form suitable for oral administration as a tablet, capsule, pill,powder, sustained release formulations, solution, and suspension. Lesspreferred (with the mesylate form being the preferred form) arecompositons for parenteral injection as a sterile solution, suspensionor emulsion, for topical administration as an ointment or cream or forrectal administration as a suppository. The pharmaceutical compositionmay be in unit dosage forms suitable for single administration ofprecise dosages. The pharmaceutical composition will include aconventional pharmaceutical carrier or excipient and a compoundaccording to the invention as an active ingredient. In addition, it mayinclude other medicinal or pharmaceutical agents, carriers, adjuvants,etc.

Exemplary parenteral administration forms include solutions orsuspensions of active compounds in sterile aqueous solutions, forexample, aqueous propylene glycol or dextrose solutions. Such dosageforms can be suitably buffered, if desired.

Suitable pharmaceutical carriers include inert diluents or fillers,water and various organic solvents. The pharmaceutical compositions may,if desired, contain additional ingredients such as flavorings, binders,excipients and the like. Thus for oral administration, tabletscontaining various excipients, such as citric acid may be employedtogether with various disintegrants such as starch, alginic acid andcertain complex silicates and with binding agents such as sucrose,gelatin and acacia. Additionally, lubricating agents such as magnesiumstearate, sodium lauryl sulfate and talc are often useful for tabletingpurposes. Solid compositions of a similar type may also be employed insoft and hard filled gelatin capsules. Preferred materials, therefor,include lactose or milk sugar and high molecular weight polyethyleneglycols. When aqueous suspensions or elixirs are desired for oraladministration the active compound therein may be combined with varioussweetening or flavoring agents, coloring matters or dyes and, ifdesired, emulsifying agents or suspending agents, together with diluentssuch as water, ethanol, propylene glycol, glycerin, or combinationsthereof. Additionally, it is also possible to administer the compound ofthe invention topically and this may be done by way of creams, jellies,gels, pastes, ointments and the like, in accordance with standardpharmaceutical practice.

The compound of the invention may also be administered to a mammal otherthan a human. The dosage to be administered to a mammal will depend onthe animal species and the disease or disorder being treated. Thecompound may be administered to animals in the form of a capsule, bolus,tablet or liquid drench. The compound may also be administered toanimals by injection or as an implant. Such formulations are prepared ina conventional manner in accordance with standard veterinary practice.As an alternative, the compound may be administered with the animalfeedstuff, and for this purpose a concentrated feed additive or premixmay be prepared for mixing with the normal animal feed.

Methods of preparing various pharmaceutical compositions with a specificamount of active compound are known, or will be apparent, to thoseskilled in this art. For examples, see Remington's PharmaceuticalSciences, Mack Publishing Company, Easter, Pa., 15th Edition (1975).

Administration and Dosage

Administration of the compounds of the present invention (hereinafterthe “active compound(s)”) can be effected by any method that enablesdelivery of the compounds to the site of action. These methodspreferably include oral routes such as in the form of tablets,intraduodenal routes, parenteral injection (including intravenous,subcutaneous, intramuscular, intravascular or infusion), topical, andrectal administration. While parenteral administration is usuallypreferred, oral administration is preferred for the hydrochloride Bpolymorph.

The amount of the active compound administered will be dependent on thesubject being treated, the severity of the disorder or condition, therate of administration and the judgement of the prescribing physician.However, an effective dosage is in the range of about 0.001 to about 100mg per kg body weight per day, preferably about 1 to about 35 mg/kg/day,in single or divided doses. For a 70 kg human, this would amount toabout 0.05 to about 7 g/day, preferably about 0.2 to about 2.5 g/day. Insome instances, dosage levels below the lower limit of the aforesaidrange may be more than adequate, while in other cases still larger dosesmay be employed without causing any harmful side effect, provided thatsuch larger doses are first divided into several small doses foradministration throughout the day.

[6,7-bis(2-methoxyethoxy)quinazolin-4-yl]-(3-ethynylphenyl)-aminehydrochloride, preferably the stable polymorph B form, at doses of1-7000 mg/day, preferably 5-2500 mg/day, most preferably 5-200 mg/day,is also useful for the treatment of patients (as measured, for example,by increased survival times) by using combination therapies, for examplein NSCLC. (IIIb/V), as a 1^(st) line therapy with carboplatin/paclitaxelor gemcitabine/cisplatin, in NSCLC (IIIb/V), as a 2^(nd) line therapywith taxotere, and in head and neck cancers, as a 2nd line therapy withmethotrexate for patients refractory to 5 FU/cisplatin.

[6,7-bis(2-methoxyethoxy)quinazolin-4-yl]-(3-ethynylphenyl)-aminehydrochloride, preferably the stable polymorph B form, at doses of1-7000 mg/day, preferably 5-2500 mg/day, most preferably 5-200 mg/day,is also useful for the treatment of patients with additional conditions,including pancreatic cancer, with or without gemcitabine co-treatment,as first line therapy, for renal cancer, gastric cancer, prostatecancer, colorectal cancer (e.g. as a 2nd line therapy for patients whohave failed 5 FU/LCV/Irinotecan therapy), and also for hepatocellular,bladder, brain, ovarian, breast, and cervical cancers. For suchtreatments, in advanced disease patients with refractory disease,treatment effectiveness is readily monitored by an increased responserate, an increased time to progression or an increase in survival time.

[6,7-bis(2-methoxyethoxy)quinazolin-4-yl]-(3-ethynylphenyl)-aminehydrochloride, preferably the stable polymorph B form, is typically usedat doses of 1-7000 mg/day, preferably 5-2500 mg/day, most preferably5-200 mg/day, for any of the above treatments.

The examples and preparations provided below further illustrate andexemplify the compounds of the present invention and methods ofpreparing such compounds. This invention will be better understood fromthe Experimental Details which follow. However, one skilled in the artwill readily appreciate that the specific methods and results discussedare merely illustrative of the invention as described more fully in theclaims which follow thereafter.

EXPERIMENTAL DETAILS Example 1 Preparation of Compound of Formula 4

The following materials were used in the synthesis of the compound offormula 4: Equivalents/ Materials Quantity Units Volumes Compound offormula 3  88.0 kg 1 equivalent Thionyl chloride  89.0 kg 2.5equivalents Dimethylformamide  11 kg 0.5 equivalent methylene chloride880.0 L 10 L/kg 50% sodium hydroxide solution as required L 1 equivalentHeptane 880.0 L 10 L/kg

The following procedure is exemplary of the procedure to follow in thesynthesis of the formula 4 compound:

-   -   88.0 kg of the compound of formula 3, 880.0 L methylene        chloride, and 11.0 kg of dimethylformamide were charged to a        clean, dry, glass-lined vessel under nitrogen atmosphere. 89 Kg        of thionyl chloride were added to the mix while it is maintained        at a temperature of a less than 30° C. during the charge. The        contents of the reaction vessel were then heated for a minimum        of five hours at reflux temperature before sampling for reaction        completion and the pH is adjusted to be maintained between 7.0        to 8.0, by using 50% NaOH, as required and the temperature of        the reaction mixture is maintained at less than 25° C. The        biphasic mixture is stirred for fifteen to twenty minutes and        allowed to settle for a minimum of thirty minutes. The layers        were separated and the organic layer was concentrated to ⅓ of        its volume by removing methylene chloride. 880 L heptane was        added with continued distillation of the remaining methylene        chloride until the distillate reaches a temperature between 65        and 68° C. The mixture was then cooled to between 10 to 15° C.        over 5 hours and granulated for a minimum of 1 hour with the        solids being isolated by filtration and washed with 220 L        heptane. The solids (formula 4 compound) were dried in a vacuum        drier at 45 to 50° C.

Example 2 Alternative Preparation of Compound of Formula 4

In the reaction shown in Example 1, sodium bicarbonate may successfullybe used instead of sodium hydroxide as shown in this Example.Equivalents/ Materials Quantity Units Volumes Compound of formula 3 30.0 kg 1 equivalent Thionyl chloride  36.4 kg 3 equivalentsDimethylformamide  3.75 kg 0.5 equivalent methylene chloride 300 L 10L/kg 50% sodium hydroxide solution as L required Heptane 375 L 12.5 L/kgHeptane (wash)  90 L 3 L/kg Sodium Bicarbonate  64.2 Kg 7.5 equivalents

-   -   30.0 kg of the compound of formula 3, 300.0L methylene chloride,        and 3.75 kg of dimethylformamide were charged to a clean, dry,        glass-lined vessel under a nitrogen atmosphere. 36.4 kg of        thionyl chloride was added to the mix while it was maintained at        a temperature of less than 30° C. during the charge. The        contents of the reaction vessel were then heated at reflux        temperature for 13 h before sampling for reaction completion.        The reaction mixture was cooled to 20-25° C. and added slowly to        a stirred solution of sodium bicarbonate 64.2 kg and water 274L        cooled to 4° C. so that the temperature was maintained at less        than 10° C. The final pH of the mixture was adjusted to within        the range 7.0 to 8.0 by using 50% sodium hydroxide solution as        required. The biphasic mixture was stirred for fifteen to twenty        minutes and allowed to settle for a minimum of thirty minutes at        10-20° C. The layers were separated and the organic layer was        concentrated to ⅓ of its volume by removing methylene chloride.        375L of heptane was added with continued distillation of the        remaining methylene chloride until the distillate reached a        temperature between 65 and 68° C. The mixture was then cooled to        0 to 5° C. over 4 hour and granulated for a minimum of 1 hour        with the solids being isolated by filtration and washed with 90L        heptane.

The solids (formula 4 compound) were dried in a vacuum drier at 45 to50° C.

Example 3 Preparation of Compound of Formulas 6 and 2 (Step 2):

The following materials were used in the synthesis of the compound offormula 6, as intermediate, and the compound of formula 2: Equivalents/Materials Quantity Units Volumes Compound of formula 5 61.1 kg 1.2equivalents Toluene 489 L 8 L/kg (WRT to formula 5 c'mpd) Sodiumhydroxide pellets 4.5 kg 0.16 equivalents Filteraid 0.5 kg 0.017 kg/kg(WRT to c'mpd 5) Compound of formula 4 90.8 kg 1.0 equivalentAcetonitrile 732 L 12 L/kg (WRT to c'mpd 5)

Example 4 Preparation of Compound of Formula 2

The following procedure is exemplary of the procedure to follow in thesynthesis of the formula 2 compound and intermediate compound of formula6:

-   -   61.1 kg of formula 5 compound, 4.5 kg sodium hydroxide pellets        and 489 L-toluene were charged to a clean, dry, reaction vessel        under nitrogen atmosphere and the reaction temperature is        adjusted to between 105 to 108° C. Acetone was removed over four        hours by atmospheric distillation while toluene is added to        maintain a minimum volume of 6 L of solvent per kg of formula 5        compound. The reaction mixture was then heated at reflux        temperature, returning distillates to pot, until the reaction        was complete. The mixture was then cooled to between 20 to 25°        C., at which time a slurry of 40.0 L toluene and 0.5 kg        filteraid was charged to the reaction mixture and the mixture        was agitated for ten to fifteen minutes. The resultant material        was filtered to remove filteraid, and the cake is washed with 30        L toluene (compound of formula 6).

The filtrate (compound of formula 6) was placed in a clean, dry reactionvessel under nitrogen atmosphere, and 90.8 kg of the compound of formula4 was charged into the reaction vessel together with 732 L acetonitrile.The reaction vessel was heated to reflux temperature and well agitated.Agitator speed was lowered when heavy solids appear. When the reactionwas complete, the contents of reaction vessel were cooled to between 19to 25° C. over three to four hours and the contents were agitated for atleast one hour at a temperature between 20 and 25° C. The solids(compound of formula 2, polymorph A form, or mixture of polymorph A andB) were then isolated by filtration and the filter cake was washed withtwo portions of 50 L acetonitrile and dried under vacuum at atemperature between 40 and 45° C.

It has been discovered that the production of the A polymorph is favoredby the reduction of the amount of acetonitrile relative to toluene, andparticularly favored if isopropanol is used in place of acetonitrile.However, the use of isopropanol or other alcohols as cosolvents isdisfavored because of the propensity to form an ether linkage betweenthe alcoholic oxygen and the 4-carbon of the quinazoline, instead of thedesired ethynyl phenyl amino moiety.

It has been further discovered that adjusting the pH of the reaction tobetween pH 1 and pH 7, preferably between pH 2 and pH 5, more preferablybetween pH 2.5 and pH 4, most preferably pH 3, will improve the rate ofthe reaction.

Example 5 Recrystallization of Compound of Formula 2 (Which may be inPolymorph A form or a Mixture of Polymorphs A and B) to Polymorph B(Step 3)

The following materials were used in the conversion of polymorph A (ormixtures of polymorphs A and B) to polymorph B of the compound offormula 2: Materials Quantity Units Equivalents/Volumes Polymorph A(formula 2) 117.6 kg 1 equivalent 2B-ethanol 1881.6 L 16 L/kg Water470.4 L 4 L/kg

The following procedure is exemplary of procedures used to convertpolymorph A (or mixtures of polymorphs A and B) into the morethermodynamically stable polymorph B of the compound of formula 2:

-   -   117.6 kg of the polymorph A (or mixtures of polymorphs A and B)        were charged to a clean, dry, reaction vessel together 1881.6 L        2B-ethanol and 470.4 L water under a nitrogen atmosphere. The        temperature was adjusted to reflux (˜80° C.) and the mixture was        agitated until the solids dissolve. The solution was cooled to        between 65 and 70° C. and clarified by filtration. With low        speed agitation, the solution was further cooled to between 50        and 60° C. over a minimum time of 2 hours and the precipitate        was granulated for 2 hours at this temperature. The mixture was        further cooled to between 0 and 5° C. over a minimum time of 4        hours and granulated for a minimum of 2 hours at this        temperature. The solids (polymorph B) were isolated by        filtration and washed with at least 100 L 2B-ethanol. The solids        were determined to be crystalline polymorph B form of        [6,7-bis(2-methoxyethoxy)quinazolin-4-yl]-(3-ethynylphenyl)-amine        hydrochloride substantially free of the polymorph A from. The        solids obtained by this method are substantially homogeneous        polymorph B form crystals relative to the polymorph A form. The        method allows for production of polymorph B in an amount at        least 70% by weight, at least 80% by weight, at least 90% by        weight, at least 95% by weight, and at least 98% by weight        relative to the weight of the polymorph A. It is to be        understood that the methods described herein are only exemplary        and are not intended to exclude variations in the above        parameters which allow the production of polymorph B in varying        granulations and yields, according to the desired storage,        handling and manufacturing applications of the compound. The        solids were vacuum dried at a temperature below 50° C. and the        resultant product was milled to provide the polymorph B in        usable form.

Example 6 Clinical Studies Utilizing Treatment with the Stable PolymorphB form of[6,7-bis(2-methoxyethoxy)quinazolin-4-yl]-(3-ethynylphenyl)-amineHydrochloride

The stable polymorph B form of[6,7-bis(2-methoxyethoxy)quinazolin-4-yl]-(3-ethynylphenyl)-aminehydrochloride is a potent, selective and orally active inhibitor of theepidermal growth factor receptor (EFGR) protein-tyrosine kinase, anoncogene that has been associated with the aberrant growth that ischaracteristic of cancer cells. This compound is being evaluated inclinical trials in normal healthy volunteers and in cancer patients inorder to assess its safety profile and effectiveness.

Phase I Clinical Studies

Phase I clinical studies of the stable polymorph B form of[6,7-bis(2-methoxyethoxy)quinazolin-4-yl]-(3-ethynylphenyl)-aminehydrochloride have been effectively completed in volunteers, initially,and subsequently in cancer patients, at single doses ranging from 25-200mg/day or 100-1600 mg/week. Data from these studies revealed no adverseevents that were greater than moderate in severity for a dose of 150mg/day. In a daily dosing regimen study the dose limiting toxicity at200 mg/day was diarrhea. This observed side effect was effectivelycontrolled at the 150 mg daily dose level using Loperamide (Imodium®).The second adverse event observed in these studies, and most significanttoxicity at 150 mg daily, was a monomorphic acneiform rash analogous tothat reported for other EGFR inhibitor agents in clinical trials. Thisrash had an “above-waist” distribution including face, scalp, neck,arms, chest and back. The rash has a unique histopathology of PMNinfiltration with mild epidermal hyperproliferation. It is notconsistent with drug hypersensitivity nor does it appear to be a “named”dermatological condition. This rash has not been a significantimpediment to patients staying on the Phase II trials. The stablepolymorph B form of[6,7-bis(2-methoxyethoxy)quinazolin-4-yl]-(3-ethynylphenyl)-aminehydrochloride has been tested in a total of 290 patients in Phase I andongoing Phase II studies and demonstrates a well tolerated safetyprofile. Furthermore, preliminary evidence of effectiveness was observedin Phase I studies. For example, in one Phase I study of 28 patients, 8patients remain alive over a year after inception of treatment and 12patients remained alive from 9-22 months.

In order to establish a suitable safety profile, the stable polymorph Bform of[6,7-bis(2-methoxyethoxy)quinazolin-4-yl]-(3-ethynylphenyl)-aminehydrochloride is also used at doses of 1-7000 mg/day, preferably 5-2500mg/day, most preferably 5-200 mg/day, in Phase I clinical combinationstudies with one or more additional drugs or treatments, preferablyselected from one of the following group—Taxol, Gemcitabine, Taxotere,Capcitabine, 5 FU, Cisplatin, Temozolomide, radiation treatment, andchemoradiation treatment.

Phase II and Phase III Clinical Studies

Three Phase II single agent studies of the stable polymorph B form of[6,7-bis(2-methoxyethoxy)quinazolin-4-yl]-(3-ethynylphenyl)-aminehydrochloride in refractory non-small cell lung cancer, advanced headand neck cancer and refractory ovarian cancer, at a 150 mg daily dosewere initiated.

Indications of single agent anti-tumor activity for the stable polymorphB form of[6,7-bis(2-methoxyethoxy)quinazolin-4-yl]-(3-ethynylphenyl)-aminehydrochloride was seen in patients with advanced cancers in severaldifferent tumor types. For example, initial findings indicate that thestable polymorph B form of[6,7-bis(2-methoxyethoxy)quinazolin-4-yl]-(3-ethynylphenyl)-aminehydrochloride is a well-tolerated oral medication that is active as amonotherapy when administered to patients with advanced head and neckcancer. In preliminary results 3 patients had objective partialresponses, while another 9 patients showed evidence of a stabilizationof their disease status. The acneiform rash, which is apparentlycharacteristic of all the anti-EGFR inhibitors undergoing clinicaltesting, was reported in approximately 70% of the first group ofpatients in this study.

The early data emerging from the 48 patient Phase II study in refractorynon-small cell lung cancer (NSCLC) patients also indicates theeffectiveness of treatment with the stable polymorph B form of[6,7-bis(2-methoxyethoxy)quinazolin-4-yl]-(3-ethynylphenyl)-aminehydrochloride as a single agent anti-tumor drug for NSCLC. Of the first19 evaluable patients in the study, 5 had objective partial responses,while another 4 patients showed evidence of a stabilization of theirdisease status. Partial responses were observed in two patients who hadbeen treated previously with two and three different chemotherapyregimens. Thus it appears that the stable polymorph B form of[6,7-bis(2-methoxyethoxy)quinazolin-4-yl]-(3-ethynylphenyl)-aminehydrochloride is a well tolerated, oral medication which is active innon-small cell lung cancer.

Qualification criteria for the open label, single agent study requiredthe patients to have failed platinum-based chemotherapy and to havetumors that are histopathologically confirmed to be EGFR positive. Theprimary endpoint in the study is response rate with stable disease andtime-to-progression amongst the secondary end-points.

Evidence of anti-tumor activity can also be seen in the patients withovarian cancer in the on-going Phase II study. In preliminary results 2patients had objective partial responses, while another 4 patientsshowed evidence of a stabilization of their disease status. Documentedevidence of anti-tumor activity was also seen in other EGFR positivetumor types, including colorectal and renal cell carcinoma, from Phase Istudies in cancer patients with multiple tumor types.

1-54. (canceled)
 55. A method of monotherapy for a subject sufferingfrom abnormal cell growth expressing the epidermal growth factorreceptor (EGFR) which comprises orally administering to the subject atherapeutically effective amount of a crystalline polymorph of thehydrochloride salt ofN-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine so as totreat the subject.
 56. The method of claim 55, wherein the abnormal cellgrowth is brain cancer, squamous cell cancer, bladder cancer, gastriccancer, pancreatic cancer, hepatic cancer, glioblastoma multiformebreast cancer, head cancer, neck cancer, esophageal cancer, prostatecancer, colorectal cancer, lung cancer, renal cancer, kidney cancer,ovarian cancer, gynecological cancer, thyroid cancer, non-small celllung cancer (NSCLC), refractory ovarian cancer, or head and neck cancer.57. The method of claim 55, wherein the abnormal cell growth isnon-small cell lung cancer (NSCLC).
 58. The method of claim 55, whereinthe abnormal cell growth is refractory ovarian cancer.
 59. The method ofclaim 55, wherein the abnormal cell growth is head and neck cancer. 60.The method of claim 55, wherein the therapeutically effective amount isfrom about 0.001 to about 100 mg/kg/day.
 61. The method of claim 55,wherein the therapeutically effective amount is from about 1 to about 35mg/kg/day.
 62. The method of claim 55, wherein the therapeuticallyeffective amount is from about 1 to about 7000 mg/day.
 63. The method ofclaim 55, wherein the therapeutically effective amount is from about 5to about 2500 mg/day.
 64. The method of claim 55, wherein thetherapeutically effective amount is from about 5 to about 200 mg/day.65. The method of claim 55, wherein the therapeutically effective amountis from about 25 to about 200 mg/day.
 66. The method of claim 55,wherein the treatment is a palliative or neo-adjuvant/adjuvantmonotherapy.
 67. The method of claim 55, wherein the EGFR receptor isthe EGFRvIII receptor.
 68. The method of claim 55, wherein thetherapeutically effective amount is from 100 to 1600 mg/week.
 69. Themethod of claim 55, wherein the therapeutically effective amount isorally administered weekly.